EPSTEIN-BARR VIRUS EXPRESSION IN NORMAL HUMAN EPITHELIUM

这项拟议研究的长期目标是识别病毒和 细胞因子，人类粘膜独有的，有助于 爱泼斯坦-巴尔病毒(EBV)的发病机制和持久性机制。 EBV相关疾病表明病毒相互作用在 人体粘膜：鼻咽癌和口腔毛状白斑。 (以EBV潜伏期和侵略性复制性感染为特征， 分别)是上皮性细胞疾病。非洲伯基特淋巴瘤， 以其不寻常的组织分布，是一种恶性肿瘤 粘膜相关淋巴组织。利用在以下方面获得的信息 目前的赠款期限，这首次表明存在 多种EBV变异株具有明显的生物学差异和组织 为了达到我们的长期目标，我们提出了以下目标：1) 裂解EBV分子结构的测定；2)评价 转化和裂解变异体在上皮和上皮细胞中的分布 粘膜系统的淋巴成分；3)机制的阐明 对于EBV在粘膜上皮成分中的持久性。 基于已知的自发实验室的生物活动 EBV核抗原2(EBNA2)编码基因缺失突变 假设相似的野生型变体代表了 我们对EBV致病机制的理解。通过分子克隆和 聚合酶链式反应分析，我们将描绘(在目标1下) 直接从咽喉取材的EBNA2缺失病毒的基因结构 洗涤，从而避免了通道进入时固有的选择偏差 淋巴细胞培养。缺失和潜在基因的证明 重排将提供生物活动改变的迹象。 在目标2中，使用适合于PCR分析的选定的基因分型标记将 允许我们通过粘膜的组织隔间追踪EBV变异体 系统，识别组织取向，并定位潜在的位置 菌株之间的相互作用。最后，我们将使用EBV毒株，同基因 除了他们的EBNA2基因，用来分析实验感染的 EB病毒可能的转化基因在上皮中的转录。在……里面 在裸鼠体内维持受感染的人类上皮细胞移植，我们将 基因表达模式与EBNA2缺失突变体能力的关系 在没有存活的B细胞的情况下维持长期感染 组件。

The long term goal of the proposed research is to identify viral and cellular factors, unique to human mucosa, which contribute to Epstein-Barr virus (EBV) pathogenesis and to mechanisms of persistence. EBV-associated diseases point to the importance of viral interactions in human mucosa: nasopharyngeal carcinoma and oral hairy leukoplakia. (characterized by EBV latency and an aggressively replicative infection, respectively) are epithelial cell diseases. African Burkitt's lymphoma, with its unusual tissue distribution, is a malignancy of the mucosal-associated lymphoid tissue. Utilizing information gained in the current grant period which suggests for the first time the existence of multiple EBV variants with distinct biologic differences and tissue tropisms, we propose the following aims to reach our long-term goals: 1) Determination of the molecular structure of lytic EBV; 2) Evaluation of the distribution of transforming and lytic variants within epithelial and lymphoid components of the mucosal system; 3) Elucidation of mechanisms for EBV persistence in the epithelial components of the mucosa. Based on the known biologic activity of a spontaneous laboratory mutant deleted for the EBV nuclear antigen 2 (EBNA2) encoding gene, we postulate that similar wild type variants represent the "missing link' in our understanding of EBV pathogenic mechanisms. By molecular cloning and polymerase chain reaction analysis, we will delineate (under aim 1) the genetic structure of EBNA2 deleted virus taken directly from throat washings, thus avoiding the selection bias inherent with passage in lymphocyte culture. Demonstration of deletions and potential gene rearrangements will provide an indication of altered biologic activity. In aim #2, use of select genotypic markers amenable to PCR analysis will allow us to track EBV variants through tissue compartments of the mucosal system, identify tissue tropisms, and locate sites for potential interaction between strains. Finally, we will use EBV strains, isogenic except for their EBNA2 gene, to analyze in experimentally infected epithelium transcription from EBV's putative transforming genes. In infected human epithelial cell implants maintained in nude mice, we will relate patterns of gene expression to the EBNA2 deletion mutant's ability to sustain long-term infection in the absence of a viable B cell component.